[1]姚言雪,董淑英,朱晨露,等.下调Pannexin2通道能增强顺铂诱导睾丸癌(I-10)细胞凋亡[J].南方医科大学学报,2020,(08):1090-1096.
 Down-regulation of pannexin 2 channel enhances cisplatin-induced apoptosis in testicularcancer I-10 cells[J].Journal of Southern Medical University,2020,(08):1090-1096.
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下调Pannexin2通道能增强顺铂诱导睾丸癌(I-10)细胞凋亡()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2020年08期
页码:
1090-1096
栏目:
出版日期:
2020-07-30

文章信息/Info

Title:
Down-regulation of pannexin 2 channel enhances cisplatin-induced apoptosis in testicular cancer I-10 cells
作者:
姚言雪董淑英朱晨露胡 淼杜宝龙童旭辉
关键词:
顺铂睾丸癌Pannexin2凋亡
Keywords:
cisplatinum testicular cancer pannexin 2 apoptosis
文献标志码:
A
摘要:
目的 探讨Pannexin2(Panx-2)蛋白在睾丸癌细胞中的表达水平以及干扰Panx-2表达对顺铂诱导睾丸癌(I-10)细胞凋亡的影响。方法 免疫印迹法Panx-2蛋白在睾丸癌细胞中的表达水平;以睾丸癌I-10细胞株为研究对象,实验分为转染试剂对照组(mork组)、阴性对照质粒组(NC组)、干扰Panx-2组(shRNA1质粒组和shRNA2质粒组),采用免疫印迹法验证Panx-2的表达;在转染细胞中添加16 μmol/L的顺铂诱导细胞死亡,通过MTT法分别检测细胞在24、48、72 h的存活率,集落克隆法检测细胞的集落形成能力;在顺铂(16 μmol/L)处理8 h后,采用AnnexinV/PI双染法检测细胞的早期凋亡;在顺铂(16 μmol/L)作用24 h后, 免疫印迹法检测细胞凋亡相关蛋白caspase 3、Bcl-2和Bax的表达水平。结果 与I-10细胞和Tcam-2细胞相比,I-10/DDP细胞(P< 0.001)和Tcam-2/DDP细胞(P<0.01)中Panx-2的表达显著增加;干扰Panx-2基因后,免疫印迹结果显示,与NC组相比,shRNA1和shRNA2组中Panx-2的表达量下降(P<0.05);MTT法显示shRNA1和shRNA2组细胞的存活率均低于NC组(P<0.001);在含顺铂培养基的培养下,集落克隆法显示shRNA1和shRNA2组细胞的集落形成能力低于NC组(P<0.001);AnnexinV/PI双染法显示shRNA1和shRNA2组的早期凋亡率(P<0.01)均低于NC组;免疫印迹结果显示,与NC组相比,shRNA1和shRNA2组中凋亡相关蛋白caspase 3表达升高(P<0.05),Bcl-2表达降低(P<0.01),Bax表达升高(P<0.05)。结论 下调Pannexin2通道能增强顺铂诱导睾丸癌(I-10)细胞凋亡。
Abstract:
Objective To investigate the effect of down-regulation of pannexin 2 (Panx-2) channels on cisplatin-induced apoptosis in I-10 cells. Methods The expression of Panx-2 protein in testicular cancer cells was detected with Western blotting. The testicular cancer cell line I-10 was transfected with two short hairpin RNA (shRNA1 and shRNA2) via Lipofectamine2000, the empty vector (NC group) or Lipofectamine2000 (blank control group), and the changes in the expression of Panx-2 was detected with Western blotting. The effects of transfection with a Panx-2 inhibitor on surviving fraction of the cells treated with cisplatin (16 μmol/L) for 24 h, 48 h and 72 h was assessed with MTT assay, and the clonogenic capacity of the cells was evaluated with colony-forming assay. At 8 h after incubation with 16 μmol/L cisplatin, AnnexinV/PI double staining was used to detect the early apoptosis of the cells. After 24 h of treatment with 16 μmol/L cisplatin, the cells were examined for expressions of caspase-3, Bcl-2 and Bax using Western blotting. Results The expression of Panx-2 was significantly increased in cisplatin-resistant I-10/DDP (P<0.001) cells and Tcam-2/DDP (P<0.01) cells as compared with I-10 cells and Tcam-2 cells. Transfection of I-10 cells with shRNA1 and shRNA2 resulted in significantly decreased Panx-2 expression (P<0.05) and significantly reduced cell surviving fraction (P<0.001). In the presence of cisplatin, the cells in NC group showed a higher clonogenic efficiency than those in shRNA1 and shRNA2 groups (P<0.001). The early-stage apoptosis rate of the cells in shRNA1 and shRNA2 groups were significantly higher than that in NC group (P<0.01). Panx-2 knockdown in I-10 cells significantly increased caspase-3 and Bax expressions (P<0.05) and significantly decreased the expression of Bcl-2 (P<0.01). Conclusion Down-regulation of Panx-2 channel enhances cisplatin-induced apoptosis in cultured testicular cancer cells.

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更新日期/Last Update: 2020-07-29